| References | 1. DabbsDavidJ. Diagnostic immunohistochemistry (M). Beijing: Peking University Medical Press, 2008.9.
2. [US] Ed Harlow, David Lane Antibody Technical Guide (M). Beijing: Science Press, 2002: 79-80, 105.
3. Stack, E.C., etc., Multiplexed immunohistochemistry, imaging, and quantitation: are view, with anassessment of Tyramide signal amplification, multispectral imaging and multiplex analysis. Methods, 2014; 70 (1): 46-58.
4. Qian Bangguo Jiao Lei Application of multi-label immunofluorescence staining and Doppler imaging technology in histological research. Chinese Journal of Histochemistry and Cytochemistry, 2017 (4): 373-382. |
| Usage | 1. Dilution of fluorescent dye: the dye is 100 × mother liquor, and the signal amplification reaction solution is diluted at 1: 100 to prepare the dye working solution (ready-to-use); DAPI prepares the working solution using sterile water dilution at 1: 100.
2. Use of secondary antibody: The kit is an anti-rabbit secondary antibody. It is not recommended that the sample be homologous to the secondary antibody. Please verify whether the species of the primary antibody matches before the experiment.
3. Frozen sections and cell crawlers must be used together with the purchase of antibody eluate (item number:abs994) |
| Theory | Multiplex fluorescence immunohistochemical staining is based on the principle of specific binding of antigen and antibody. The secondary antibody labeled with horseradish peroxidase (HRP) is selected to activate the fluorescent dye in the kit and covalently bind the signal to the antigen. In situ multi-target staining of tissues or cells is achieved by multiple rounds of staining cycles with the aid of different fluorescent dye labels. |
| Synonym | Multicolor kit |
| Description | There are complex cells in tissue microenvironment, and the phenotype, state, abundance and distribution of these cells have important biological significance and clinical value. It can be rendered in situ in tissue by means of antibody staining. Immunohistochemical staining is a common technique to study tissue morphology and protein expression in situ. Routine IHC detection can only show a single index, and it is difficult to present the cell composition, state and relationship in the complex tissue microenvironment. This information is crucial to the diagnosis and treatment of diseases! Principle of tyrosine signal amplification technology: Similar to the DAB color development method of conventional immunohistochemistry, TSA technology also uses HRP-labeled secondary antibody. HRP catalyzes the fluorescein substrate added to the system to produce an activated fluorescein substrate, which can be covalently bound to the antigen. Tyrosine is covalently bound to stably covalently bound to fluorescein on the sample. After that, the non-covalently bound antibody is washed away by thermal repair method, replaced with a primary antibody for the second round of incubation, and replaced with another fluorescein substrate, so that multiple labeling can be achieved by reciprocating. |
| Composition | TSA monochromatic fluorescent dye 520, TSA monochromatic fluorescent dye 650, signal amplification reaction solution, anti-rabbit HRP labeled secondary antibody, anti-fluorescence quenching sealing tablet, DAPI |
| General Notes | 1. This kit is only used for immunohistochemistry and is not used for other purposes.
2. This kit is restricted to professional use only.
3. Appropriate protective measures should be taken to avoid contact of the reagent with the skin and eyes.
4. The activity of reagents beyond the expiration date may be reduced, so reagents beyond the expiration date should not be used.
5. If the dyeing components of this kit are mixed with products of other companies, abnormalities may occur during the dyeing process.
6. Incomplete dewaxing will easily affect the dyeing effect.
7. In order to prevent possible false negative and false positive results, positive control and negative control should be carried out at the same time during the experiment.
8. All kinds of wastes generated in the use of this kit should be disposed of in accordance with the Regulations on Medical Waste Management. |
| Storage Temp. | Fluorescent dyes should be stored in the dark from light at 2-8 ℃, and the validity period is 12 months from receipt of the goods. |
| Applications | It is mainly used for scientific research on immunohistochemical staining of tissues, paraffin sections and TMA chips, and cannot be used for in vitro clinical diagnosis or human experiments. |
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